Name: selective viral transduction of adult-born olfactory neurons for chronic in vivo optogenetic stimulation
Uploaded: 2011-12-28T12:00:00
Description: Watch this Scientific Journal Video about Selective Viral Transduction of Adult-born Olfactory Neurons for Chronic in vivo Optogenetic Stimulation at JoVE.com

Questo lavoro è stato sostenuto dalla compagnia di assicurazione sulla vita &quot;Novalis-Taitbout&quot;, Ecole des Neurosciences de Paris (PEV), l&#39;Agence Nationale de la Recherche &quot;ANR-09-Neur-004&quot;, nella cornice di &quot;NEURON ERA-NET&quot; di 7 ° PQ dalla Commissione Europea, &quot;Fondazione pour la Recherche Medicale&quot;, la Fondazione Letten e la Fondazione Pasteur.

1. Stereotassica iniezioni <ol><li> Il virus usato in questi esperimenti è una replica con deficit di vettori lentivirali basati sul virus HIV per esprimere Channelrhodopsine2-YFP (gialla proteina fluorescente) fusione costruire guidata da un promotore sinapsina 6. Il plasmide è stato generosamente forniti da K. Deisseroth &#39;s di ricerca del gruppo ( <a href="http://www.stanford.edu/group/dlab/optogenetics/index.html">http://www.stanford.edu/group/dlab/optogenetics/index.html</a> ). Lentivirus possono ...

<ol>
	<li>Lledo, P. M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Adult+neurogenesis+and+functional+plasticity+in+neuronal+circuits.">Adult neurogenesis and functional plasticity in neuronal circuits.</a> Nat. Rev. Neurosci. 7, 179-193 (2006).</li><li>Alonso, M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Turning+astrocytes+from+the+rostral+migratory+stream+into+neurons:+a+role+for+the+olfactory+sensory+organ.">Turning astrocytes from the rostral migratory stream into neurons: a role for the olfactory sensory organ.</a> J. 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A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Targeted+optogenetic+stimulation+and+recording+of+neurons+in+vivo+using+cell-type-specific+expression+of+Channelrhodopsin-2.">Targeted optogenetic stimulation and recording of neurons in vivo using cell-type-specific expression of Channelrhodopsin-2.</a> Nature protocols. 5, 247-254 (2010).</li><li>Huber, D. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Sparse+optical+microstimulation+in+barrel+cortex+drives+learned+behaviour+in+freely+moving+mice.">Sparse optical microstimulation in barrel cortex drives learned behaviour in freely moving mice.</a> Nature. 451, 61-64 (2010).</li><li>Arenkiel, B. R. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=In+vivo+light-induced+activation+of+neural+circuitry+in+transgenic+mice+expressing+channelrhodopsin-2.">In vivo light-induced activation of neural circuitry in transgenic mice expressing channelrhodopsin-2.</a> Neuron. 54, 205-218 .</li><li>Albeanu, D. F. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=LED+arrays+as+cost+effective+and+efficient+light+sources+for+widefield+microscopy.">LED arrays as cost effective and efficient light sources for widefield microscopy.</a> PLoS ONE. 3, 2146-2146 (2008).</li><li>Slotnick, B., Bodyak, N. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Odor+Discrimination+and+Odor+Quality+Perception+in+Rats+with+Disruption+of+Connections+between+the+Olfactory+Epithelium+and+Olfactory+Bulbs.">Odor Discrimination and Odor Quality Perception in Rats with Disruption of Connections between the Olfactory Epithelium and Olfactory Bulbs.</a> J. 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Strumenti Optogenetic hanno recentemente aumentato il nostro controllo sulle popolazioni di neuroni selettivi, tra cui i neuroni adulti nati nel sistema olfattivo. Qui si descrivono come eseguire una iniezione stereotassica di un vettore lentivirus che esprimono ChR2-YFP in una specifica popolazione di adulti nati neuroni. Con un attento monitoraggio del periodo post-iniezione, siamo in grado di analizzare e manipolare l&#39;attività di una coorte di adulti nati i neuroni con un età relativa omogenea. <p class="jo...

<table border="1"><tbody><tr><td> Materiale </td><td> Azienda </td><td> Catalogo N. </td></tr><tr><td> Ketamina </td><td> Merial </td><td> Imalgène 1000 </td></tr><tr><td> Xylazina </td><td> Bayer Health Care </td><td> Rompum 0,2% </td></tr><tr><td> Pipetta Puller </td><td> Sutter </td><td> P-97 </td></tr><tr><td> Capillari di vetro (3,5 &quot;) </td><td> Drummond scientifico </td><td> 3-000-203-G / X </td></tr><tr><td> Nanoject II Microinjector </td><td> Drummond scientifico </td><td> 3-000-204 </td></tr><tr><td> Trapano ad alta chirurgia </td><td> Strumenti Scienza multa </td><td> 18000-17 </td></tr><tr><td> Micro Drill acciaio Frese punta 0,7 millimetri di diametro </td><td> Strumenti Scienza multa </td><td> 19008-07 </td></tr><tr><td> CMS LED blu 4,6 W </td><td> OSRAM </td><td> LBW5SN </td></tr><tr><td> Femmina connettore elettrico (2,54 mm) </td><td> Fischer Elektronik </td><td> BL5.36Z </td></tr><tr><td> Regolatore di corrente per LED (max: 1A) </td><td> A1W Electronik </td><td> HKO-KL50-1000 </td></tr><tr><td> Generatore di impulsi </td><td> AMPI </td><td> Maestro-8 </td></tr><tr><td> Misuratore di potenza ottica </td><td> Thor Labs </td><td> PM100 </td></tr><tr><td> Cianoacrilato </td><td> Loctite </td><td> 454 </td></tr><tr><td> Policarbossilato cemento (cemento primo strato) </td><td> Septodont </td><td> Selfast (# 0068Q) </td></tr><tr><td> Metacrilato di cemento (cemento secondo strato) </td><td> GC InternationalCorp. </td><td> Unifast Trad - liquidi (# 339292) Unifast Trad - Polvere (# 339114) </td></tr><tr><td> Carprofene </td><td> Pfizer </td><td> Rimadyl </td></tr></tbody></table>

selective viral transduction of adult-born olfactory neurons for chronic in vivo optogenetic stimulation

Interneuroni locali sono continuamente rigenerati nel bulbo olfattivo di roditori adulti 1-3. In questo processo, chiamato neurogenesi adulta, le cellule staminali neurali nelle pareti del ventricolo laterale danno luogo a neuroblasti che migrano per diversi millimetri lungo il flusso migratorio rostrale (RMS) per raggiungere e incorporare nel bulbo olfattivo. Per studiare le diverse fasi e l&#39;impatto di adulti nati integrazione dei neuroni preesistenti in circuiti olfattiva, è necessario etichettare in modo selettivo e manipolare l&#39;attività di questa specifica popolazione di neuroni. Il recente sviluppo delle tecnologie optogenetic offre l&#39;opportunità di usare la luce per attivare appunto questo gruppo specifico di neuroni senza intaccare i neuroni circostanti 4,5. Qui vi presentiamo una serie di procedure per viralmente esprimere Channelrhodopsin2 (ChR2)-YFP in una coorte temporalmente ristretto di neuroblasti in RMS prima di raggiungere il bulbo olfattivo e diventare adulto-nato neurons. Inoltre, ci mostrano come impianto e calibrare una miniatura a LED per la stimolazione cronica in vivo di ChR2-esprimere i neuroni.

Watch this Scientific Journal Video about Selective Viral Transduction of Adult-born Olfactory Neurons for Chronic in vivo Optogenetic Stimulation at JoVE.com

Selective Viral Transduction of Adult-born Olfactory Neurons for Chronic in vivo Optogenetic Stimulation

I neuroni adulti, nato del bulbo olfattivo può essere controllato tramite optogenetically Channelrhodopsin2 che esprimono iniezione lentivirali nel flusso migratorio rostrale e fotostimolazione cronica con una miniatura impiantato LED.

Selettiva trasduzione virale di neuroni olfattivi per adulti nati per cronica In vivo Optogenetic Stimolazione

Local interneurons are continuously regenerated in the olfactory bulb of adult rodents1-3. In this process, called adult neurogenesis, neural stem cells ...

Research

JoVE Journal

Neuroscience

Selective Viral Transduction of Adult-born Olfactory Neurons for Chronic in vivo Optogenetic Stimulation

Studying the Integration of Adult-born Neurons

Studiare l&#39;integrazione degli adulti nati Neuroni

Neurogenesis occurs in adult mammalian brains in the sub-ventricular zone (SVZ) of the lateral ventricle and in the sub-granular zone (SGZ) of the ...

Ricerca

Neuroscienze

Lentivirus-mediated Genetic Manipulation and Visualization of Olfactory Sensory Neurons in vivo

Lentivirus-mediated Genetic Manipulation and Visualization of Olfactory Sensory Neurons in vivo

Lentivirus-mediata manipolazione genetica e la visualizzazione dei neuroni sensoriali olfattivi In vivo</em

Development of a precise olfactory circuit relies on accurate projection of olfactory sensory neuron (OSN) axons to their synaptic 
targets in the ...

Flash Photolysis of Caged Compounds in the Cilia of Olfactory Sensory  Neurons

Fotolisi lampo di composti Caged nel Cilia di neuroni sensoriali olfattivi

Photolysis of caged compounds allows the production of rapid and localized increases in the concentration of various physiologically active compounds1. ...

Fiber-optic Implantation for Chronic Optogenetic Stimulation of Brain Tissue

Fibra ottica Impianto per la Stimolazione Optogenetic cronica del tessuto cerebrale

Elucidating patterns of neuronal connectivity has been a challenge for both clinical and basic neuroscience. Electrophysiology has been the gold standard ...

Optogenetic Stimulation of Escape Behavior in Drosophila melanogaster

Optogenetic Stimulation of Escape Behavior in Drosophila melanogaster

Stimolazione Optogenetic del comportamento in Fuga<em&gt; Drosophila melanogaster</em

A  growing number of genetically encoded tools are becoming available that allow non-invasive  manipulation of the neural activity of specific neurons in ...

A Method for High Fidelity Optogenetic Control of Individual Pyramidal Neurons In vivo

A Method for High Fidelity Optogenetic Control of Individual Pyramidal Neurons In vivo

Un metodo per High Fidelity optogenetic controllo dei singoli Piramidale neuroni<em&gt; In vivo</em

Optogenetic methods  have emerged as a powerful tool for elucidating neural circuit activity  underlying a diverse set of behaviors across a broad range ...

In vivo Optogenetic Stimulation of the Rodent Central Nervous System

In vivo Optogenetic Stimulation of the Rodent Central Nervous System

In vivo optogenetic stimolazione del sistema nervoso centrale Roditore

The ability to probe defined neural circuits in awake, freely-moving animals with cell-type specificity, spatial precision, and high temporal resolution ...

Intracerebroventricular Viral Injection of the Neonatal Mouse Brain for Persistent and Widespread Neuronal Transduction

Intracerebroventricular Iniezione virale del cervello neonatale mouse per persistente e diffusa trasduzione neuronale

With the pace of scientific advancement accelerating rapidly, new methods are needed for experimental neuroscience to quickly and easily manipulate gene ...

Optogenetic Stimulation of the Auditory Nerve

Stimolazione optogenetic del nervo uditivo

Direct electrical stimulation of spiral ganglion neurons (SGNs) by cochlear implants (CIs) enables open speech comprehension in the majority of implanted ...

Ex Vivo Optogenetic Dissection of Fear Circuits in Brain Slices

Ex Vivo Optogenetic Dissection of Fear Circuits in Brain Slices

Ex Vivo optogenetic dissezione di circuiti paura nel cervello fette

Optogenetic approaches are now widely used to study the function of neural populations and circuits by combining targeted expression of light-activated ...